Additional benefit of trastuzumab was revealed as a primary treatment for advanced gastric cancer with HER2-positive by ToGA trial. The treatment of second-line treatment for patients with primary refractory is unknown, however. There are many questions, such as seroconverted during the course of treatment of false-negative cases. It can be expected that the earliest second-line HER2-positive gastric cancer is Lapatinib, a tyrosine kinase inhibitor of both HER2 and EGFR. On the other hand, numerous clinical trials related to ‘Beyond Progression’ of additional benefit for continuing trastuzumab is conducting in many Oncology Group. HER2-positive gastric cancer is limited and coordination with each group efficiently is needed. In conducting clinical trials, it would be more important to manage the collection and analysis of serum and tissues samples for the purpose of biomarker study.

We describe the detoured migration route of the Oriental honey-buzzard Pernis ptilorhyncus, showing differences between autumn and spring migration, using data from 10 adult individuals marked with satellite transmitters. In autumn, the migration routes were very similar from Japan to the south end of the Malay Peninsula. The wintering sites were distributed within the Philippines, Borneo and the Malay Archipelago. During autumn, migration of the birds had few long-term stopover sites, instead, sometimes decidedly slowing their migration rate while proceeding in a consistent direction. During spring migration, the honey-buzzards penetrated into southern China, moving north to the base of the Korean Peninsula. The birds then went south through the Korean Peninsula to reach Japan. Before travelling to China, all spring migrants stopped for several weeks in south-east Asia. The slow rate of travel in the autumn suggests that migrants were foraging and replenishing their energy reserves. Instead of a migration strategy that uses only a few long-term stopover sites, honey-buzzards may adopt a strategy based on a number of short-term stay sites.

Legged locomotion of the Eurasian nuthatch Sitta europaea on horizontal and vertical substrates was examined using field observations and experiments. Although previous studies have reported that nuthatches use 'walking' on vertical substrates, we found that they usually used 'hopping' on both vertical and horizontal substrates. When climbing up a vertical Substrate, the feet were staggered in position and small phase differences were observed between the left and right leg movements in the gait. In upward climbing, the body was inclined towards the substrate during the first stance phase similar to other tree-trunk climbers, but the tail was not used for helping body rotation unlike most tree-trunk climbers. The staggered position of the feet may allow the legs to play different roles in pulling towards and pushing away from the substrate. In downward climbing, the feet were staggered in position, but the phase difference was quite small. In field observations, the Eurasian nuthatch preferred to move vertically, rather than in an inclined direction.

Single-molecule imaging of the entrance of a protein into the hydrophobic environment of a cell membrane was investigated. The pre-stem of LukF, one of the two components of the pore-forming toxin staphylococcal gamma-hemolysin, was specifically labeled with 6-bromoacetyl-2-dimethylaminonaphthalene (Badan), an environment-sensitive fluorophore. Incubation of this derivative with erythrocyte ghost membranes resulted in a pronounced increase in fluorescence indicating insertion of Badan into the hydrophobic interior of the lipid bilayers. However, the increase in fluorescence was completely dependent on the interaction of Badan-labeled LukF with the gamma-hemolysin second component. Individual spots of Badan fluorescence on erythrocyte membranes were visualized that were associated with single pores. Analyses of the intensities of these fluorescent spots and their photobleaching independently showed that a single pore contained 3-4 LukF molecules. Thus, environment-sensitive fluorophore signals can be used to study the insertion of specific protein domains into cell membranes at the single-molecule lever, and the use of this approach in the present study revealed that a single gamma-hemolysin pore opening contains at least three LukF molecules.

This study investigated the costs and benefits of flocking in white-fronted geese Anser albifrons foraging on rice grains in Japan. The time budgets of focal geese were recorded, and the effects of flock size on the proportions of time spent in vigilant and agonistic behaviour were tested. The results showed that the decline in vigilance level and consequent increase in foraging time were beneficial results of flocking whereas agonistic interactions, a potential cost of flocking, did not increase with increasing flock size. However, seasonal variation in flock size suggested that exploitative competition could be a cost of flocking; the sizes of flocks in spring, when resource depletion had progressed, were significantly reduced compared with those in autumn. An experimental increase in rice density resulted in a significant increase in flock size. We conclude that the flock size of foraging white-fronted geese is a result of compromise between a constant benefit of flocking (i.e. decline in vigilance level) and a cost of flocking varying with food abundance (i.e. exploitative competition).

Structural differences between dynein and kinesin suggest a unique molecular mechanism of dynein motility. Measuring the mechanical properties of a single molecule of dynein is crucial for revealing the mechanisms underlying its movement. We measured the step size and force produced by single molecules of active cytoplasmic dynein by using an optical trap and fluorescence imaging with a high temporal resolution. The velocity of dynein movement, 800 nm/s, is consistent with that reported in cells. The maximum force of 7-8 pN was independent of the ATP concentration and similar to that of kinesin. Dynein exhibited forward and occasional backwards steps of approximately 8 nm, independent of load. It is suggested that the large dynein heads take 16-nm steps by using an overlapping hand-over-hand mechanism.

Few studies have compared the relative importance of stand-level variables and landscape-level variables for wildlife in low-contrast landscapes in winter. We examined the influence of stand and context variables on bird communities in a rural landscape in Japan. Line transect surveys were conducted during the 2001 wintering season in 25 coppice and 18 conifer plantation stands located in Tochigi Prefecture, central Japan. Bird species were assigned to five guilds based on their autoecology. The relationships between the presence/absence of each guild and environmental variables were explored using logistic regression analyses. The main variables associated with the presence of litter-searchers were stand variables, while the presence of seed-eaters was mainly explained by context variables. The presence of shrub-users, canopy-users, and edge groups were explained by both variables.

Cholangiocarcinomas are usually fatal neoplasms originating from bile duct epithelia. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising agent for cancer therapy, including cholangiocarcinoma. However, many cholangiocarcinoma cells are resistant to TRAIL-mediated apoptosis. Thus, our aim was to examine the intracellular mechanisms responsible for TRAIL resistance in human cholangiocarcinoma cell lines. Three TRAIL-resistant human cholangiocarcinoma cell lines were identified. All of the cell lines expressed TRAIL receptor 1/death receptor 4 (TRAIL-R1/DR4) and TRAIL-R2/DR5. Expression of TRAIL decoy receptors and the antiapoptotic cellular FLICE-inhibitory protein (cFLIP) was inconsistent across the cell lines. Of the antiapoptotic Bcl-2 family of proteins profiled (Bcl-2, Bcl-xL, and Mcl-1), Mcl-1 was uniquely overexpressed by the cell lines. When small-interfering-RNA (siRNA) technology was used to knock down expression of Bcl-2, Bcl-xL, and Mcl-1, only the Mcl-1-siRNA sensitized the cells to TRAIL-mediated apoptosis. In a cell line stably transfected with Mcl-1-small-hairpin-RNA (Mcl-1-shRNA), Mcl-1 depletion sensitized cells to TRAIL-mediated apoptosis despite Bcl-2 expression. TRAIL-mediated apoptosis in the stably transfected cells was associated with mitochondrial depolarization, Bax activation, cytochrome c release from mitochondria, and caspase activation. Finally, flavopiridol, an anticancer drug that rapidly down-regulates Mcl-1, also sensitized cells to TRAIL cytotoxicity. In conclusion, these studies not only demonstrate that Mcl-1 mediates TRAIL resistance in cholangiocarcinoma cells by blocking the mitochondrial pathway of cell death but also identify two strategies for circumventing this resistance.

We investigated the application of satellite tracking to the conservation of cranes and other waterbirds and the necessity of international cooperation in the conservation of migratory species. Using satellite tracking, we followed 11 White-naped Cranes (Grus vipio) on migration from their breeding grounds in eastern Russia to their wintering grounds in China and Japan. From 1991 to 1993, we captured cranes with the aid of helicopters and attached satellite transmitters (platform transmitter terminals) to captured birds via a harness system. We tracked cranes for 156 days on average, across 2558 km, and obtained an average of 339 locations per crane. Cranes migrated over 8-90 days. During migration, birds spent 1-30 days at 4-12 rest sites. Cranes wintered at two main sites: Poyang Lake, China, and Izumi, Japan. All seven cranes migrating to the Poyang Lake area rested at the Yellow River delta-Bohai Bay, China, and all three traveling to Izumi rested in the Demilitarized Zone on the Korean Peninsula. Other important rest sites were marshes around the Wulagai River, Huainan, Hu-Lun Lake, Linyi, Tangshan, the Three Rivers (Sanjiang) Plain, Tianjin, and the Xar Moron River, all in China, and Lake Khanka-Xinghai at the border of China and Russia. The habitats resting cranes used most frequently were plains, including upland areas, marshes, and rivers. Although nature reserves exist at the two main wintering sites of Poyang Lake and Izumi, rest sites used by cranes were poorly protected. Even when areas used by cranes for resting or wintering were included in nature reserves, reserves were threatened by human encroachment and development. To ensure that cranes can continue to migrate successfully, it is crucial that the establishment of reserves continues at important rest areas and that the areas covered by reserves at wintering sites be extended to include more of the areas utilized by cranes. Also, development and human disturbance should be minimized in reserve areas. Because long-distance migrant birds, including cranes, range over such large areas, conservation of these organisms and their habitats necessitates multinational communication and cooperation.

Acute ethanol exposure induces oxidative stress and apoptosis in primary rat hepatocytes. Previous data indicate that the mitochondrial permeability transition (MPT) is essential for ethanol-induced apoptosis. However, the mechanism by which ethanol induces the MPT remains unclear. In this study, we investigated the role of Bax, a proapoptotic Bcl-2 family protein, in acute ethanol-induced hepatocyte apoptosis. We found that Bax translocates from the cytosol to mitochondria before mitochondrial cytochrome c release. Bax translocation was oxidative stress dependent. Mitochondrial Bax formed a protein complex with the mitochondrial voltage-dependent anion channel (VDAC). Prevention of Bax-VDAC interactions by a microinjection of anti-VDAC antibody effectively prevented hepatocyte apoptosis by ethanol. In conclusion, these data suggest that Bax translocation from the cytosol to mitochondria leads to the subsequent formation of a Bax-VDAC complex that plays a crucial role in acute ethanol-induced hepatocyte apoptosis.

Intracellular rheology is a useful probe of the mechanisms underlying spontaneous or chemotactic locomotion and transcellular migration of leukocytes. We characterized regional rheological differences between the leading, body, and trailing regions of isolated, adherent, and spontaneously locomoting human neutrophils. We optically trapped intracellular granules and measured their displacement for 500 ms after a 100-nm step change in the trap position. Results were analyzed in terms of simple viscoelasticity and with the use of structural damping (stress relaxation follows a power law in time). Structural damping fit the data better than did viscoelasticity. Regional viscoelastic stiffness and viscosity or structural damping storage and loss moduli were all significantly lower in leading regions than in pooled body and/or trailing regions (the latter were not significantly different). Structural damping showed similar levels of elastic and dissipative stresses in body and/or trailing regions; leading regions were significantly more fluidlike (increased power law exponent). Cytoskeletal disruption with cytochalasin D or nocodazole made body and/or trailing regions approximately 50% less elastic and less viscous. Cytochalasin D completely suppressed pseudopodial formation and locomotion; nocodazole had no effect on leading regions. Neither drug changed the dissipation-storage energy ratio. These results differ from those of studies of neutrophils and other cell types probed at the cell membrane via beta(2)-integrin receptors, which suggests a distinct role for the cell cortex or focal adhesion complexes. We conclude that 1) structural damping well describes intracellular rheology, and 2) while not conclusive, the significantly more fluidlike behavior of the leading edge supports the idea that intracellular pressure may be the origin of motive force in neutrophil locomotion.